RESUMEN
KEY MESSAGE: In this work, we genetically characterized the function of Arabidopsis thaliana, LONGIFOLIA (LNG1), LNG2, LNG3, LNG4, their contribution to regulate vegetative architecture in plant. We used molecular and biophysical approaches to elucidate a gene function that regulates vegetative architecture, as revealed by the leaf phenotype and later effects on flowering patterns in Arabidopsis loss-of-function mutants. As a result, LNG genes play an important role in polar cell elongation by turgor pressure controlling the activation of XTH17 and XTH24. Plant vegetative architecture is related to important traits that later influence the floral architecture involved in seed production. Leaf morphology is the primary key trait to compose plant vegetative architecture. However, molecular mechanism on leaf shape determination is not fully understood even in the model plant A. thaliana. We previously showed that LONGIFOLIA (LNG1) and LONGIFOLIA2 (LNG2) genes regulate leaf morphology by promoting longitudinal cell elongation in Arabidopsis. In this study, we further characterized two homologs of LNG1, LNG3, and LNG4, using genetic, biophysical, and molecular approaches. Single loss-of-function mutants, lng3 and lng4, do not show any phenotypic difference, but mutants of lng quadruple (lngq), and lng1/2/3 and lng1/2/4 triples, display reduced leaf length, compared to wild type. Using the paradermal analysis, we conclude that the reduced leaf size of lngq is due to decreased cell elongation in the direction of longitudinal leaf growth, and not decreased cell proliferation. This data indicate that LNG1/2/3/4 are functionally redundant, and are involved in polar cell elongation in Arabidopsis leaf. Using a biophysical approach, we show that the LNGs contribute to maintain high turgor pressure, thus regulating turgor pressure-dependent polar cell elongation. In addition, gene expression analysis showed that LNGs positively regulate the expression of the cell wall modifying enzyme encoded by a multi-gene family, xyloglucan endotransglucosylase/hydrolase (XTH). Taking all of these together, we propose that LNG related genes play an important role in polar cell elongation by changing turgor pressure and controlling the activation of XTH17 and XTH24.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/genética , Genes de Plantas , Glicosiltransferasas/metabolismo , Células Vegetales/metabolismo , Arabidopsis/citología , Arabidopsis/crecimiento & desarrollo , Proteínas de Arabidopsis/genética , Flores , Regulación de la Expresión Génica de las Plantas , Glicosiltransferasas/genética , Mutación , Hojas de la Planta/genética , Hojas de la Planta/crecimiento & desarrolloRESUMEN
Hydrogen peroxide (H 2O 2) is a reactive oxygen species that signals between cells, and H 2O 2 signaling is essential for diverse cellular processes, including stress response, defense against pathogens, and the regulation of programmed cell death in plants. Although plasma membrane intrinsic proteins (PIPs) have been known to transport H 2O 2 across cell membranes, the permeability of each family member of PIPs toward H 2O 2 has not yet been determined in most plant species. In a recent study, we showed that certain isoforms of Arabidopsis thaliana AtPIPs, including AtPIP2;2, AtPIP2;4, AtPIP2;5, and AtPIP2;7, are permeable for H 2O 2 in yeast cells. Since the expression of PIPs is differently modulated in Arabidopsis by abiotic stress or H 2O 2 treatment, it is important to investigate the integrated regulation of aquaporin expression and their physiological significance in H 2O 2 transport and plant response to diverse abiotic stresses.
Asunto(s)
Adaptación Fisiológica , Acuaporinas/metabolismo , Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Membrana Celular , Peróxido de Hidrógeno/metabolismo , Estrés Fisiológico , Adaptación Fisiológica/genética , Acuaporinas/genética , Arabidopsis/genética , Arabidopsis/fisiología , Proteínas de Arabidopsis/genética , Transporte Biológico , Membrana Celular/metabolismo , Expresión Génica , Regulación de la Expresión Génica de las Plantas , Genes de Plantas , Permeabilidad , Isoformas de Proteínas , Transducción de Señal , Estrés Fisiológico/genética , Levaduras/metabolismoRESUMEN
Although aquaporins have been known to transport hydrogen peroxide (H(2)O(2)) across cell membranes, the H(2)O(2)-regulated expression patterns and the permeability of every family member of the plasma membrane intrinsic protein (PIP) toward H(2)O(2) have not been determined. This study investigates the H(2)O(2)-regulated expression levels of all plasma membrane aquaporins of Arabidopsis thaliana (AtPIPs), and determines the permeability of every AtPIP for H(2)O(2) in yeast. Hydrogen peroxide treatment of Arabidopsis down-regulated the expression of AtPIP2 subfamily in roots but not in leaves, whereas the expression of AtPIP1 subfamily was not affected by H(2)O(2) treatment. The growth and survival of yeast cells that expressed AtPIP2;2, AtPIP2;4, AtPIP2;5, or AtPIP2;7 was reduced in the presence of H(2)O(2), while the growth of yeast cells expressing any other AtPIP family member was not affected by H(2)O(2). These results show that only certain isoforms of AtPIPs whose expression is regulated by H(2)O(2) treatment are permeable for H(2)O(2) in yeast cells, and suggest that the integrated regulation of aquaporin expression by H(2)O(2) and the capacity of individual aquaporin to transport H(2)O(2) are important for plant response to H(2)O(2).
Asunto(s)
Acuaporinas/metabolismo , Proteínas de Arabidopsis/metabolismo , Arabidopsis/citología , Arabidopsis/metabolismo , Permeabilidad de la Membrana Celular , Membrana Celular/metabolismo , Peróxido de Hidrógeno/metabolismo , Acuaporinas/antagonistas & inhibidores , Acuaporinas/genética , Arabidopsis/efectos de los fármacos , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Membrana Celular/efectos de los fármacos , Permeabilidad de la Membrana Celular/efectos de los fármacos , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Peróxido de Hidrógeno/farmacología , Viabilidad Microbiana/efectos de los fármacos , Hojas de la Planta/efectos de los fármacos , Hojas de la Planta/genética , Raíces de Plantas/efectos de los fármacos , Raíces de Plantas/genética , Saccharomyces cerevisiae/citología , Saccharomyces cerevisiae/efectos de los fármacos , Saccharomyces cerevisiae/crecimiento & desarrollo , Saccharomyces cerevisiae/metabolismo , Nitrato de Plata/farmacologíaRESUMEN
Although the number of reports demonstrating the roles of individual aquaporins in plants under diverse physiological conditions is expanding, the importance of interactions between different aquaporin isoforms and their integrated functions under stress conditions remain unclear. Here, we expressed one cucumber aquaporin gene, designated CsPIP1;1, and one figleaf gourd aquaporin gene, designated CfPIP2;1, in Arabidopsis thaliana, and investigated the effect of its expression on the natural expression patterns of endogenous PIP genes under stress conditions. The transcript levels of endogenous Arabidopsis PIP members were altered differently depending on stress conditions by the expression of CsPIP1;1 or CfPIP2;1. The transgenic Arabidopsis plants that constitutively express CfPIP2;1 displayed better growth compared with the wild-type plants under dehydration stress conditions, whereas CsPIP1;1 expression exerted a negative effect on the growth of Arabidopsis under dehydration stress conditions. CsPIP1;1 or CfPIP2;1 expression facilitated seed germination under high salt stress conditions, but had no influence on the growth of Arabidopsis under cold stress conditions. Our results indicate that the ectopic expression of a foreign aquaporin gene perturbs differently the natural expression patterns of endogenous aquaporin genes depending on particular stress conditions, and thereby influences the responses of plants to different stress conditions. This implies that the up- and/or down-regulation of aquaporins and their integrated functions are crucial to the maintenance of proper water balance under stress conditions.
Asunto(s)
Acuaporinas/genética , Proteínas de Arabidopsis/genética , Arabidopsis/genética , Regulación de la Expresión Génica de las Plantas , Acuaporinas/metabolismo , Arabidopsis/crecimiento & desarrollo , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Frío , Cucumis sativus/genética , Cucurbita/genética , Deshidratación , Genes de Plantas , Plantas Modificadas Genéticamente , Cloruro de Sodio/farmacologíaRESUMEN
Despite the high isoform multiplicity of aquaporins in plants, with 35 homologues including 13 plasma membrane intrinsic proteins (PIPs) in Arabidosis thaliana, the individual and integrated functions of aquaporins under various physiological conditions remain unclear. To better understand aquaporin functions in plants under various stress conditions, we examined transgenic Arabidopsis and tobacco plants that constitutively overexpress Arabidopsis PIP1;4 or PIP2;5 under various abiotic stress conditions. No significant differences in growth rates and water transport were found between the transgenic and wild-type plants when grown under favorable growth conditions. The transgenic plants overexpressing PIP1;4 or PIP2;5 displayed a rapid water loss under dehydration stress, which resulted in retarded germination and seedling growth under drought stress. In contrast, the transgenic plants overexpressing PIP1;4 or PIP2;5 showed enhanced water flow and facilitated germination under cold stress. The expression of several PIPs was noticeably affected by the overexpression of PIP1;4 or PIP2;5 in Arabidopsis under dehydration stress, suggesting that the expression of one aquaporin isoform influences the expression levels of other aquaporins under stress conditions. Taken together, our results demonstrate that overexpression of an aquaporin affects the expression of endogenous aquaporin genes and thereby impacts on seed germination, seedling growth, and stress responses of the plants under various stress conditions.